Our long term goal is to elucidate the mechanisms of initiation and[unreadable] regulation of RNA polymerase II transcription. Our experimental approach[unreadable] is to reconstitute these processes with purified proteins from yeast, and[unreadable] bring the power of combined biochemical and genetic analysis to bear on[unreadable] the mechanisms. During the previous project period, we developed a fully[unreadable] defined, appropriately regulated transcription system. The sub cloning of[unreadable] genes for all components and expression of many of the proteins in[unreadable] recombinant form should enable the culmination of our efforts during the[unreadable] coming project period. Specific aims of the proposed research are as[unreadable] follows:[unreadable] [unreadable] [unreadable] 1. Reconstitution and functional analysis of TFIIH.[unreadable] [unreadable] 2. Reconstitution and functional analysis of mediator.[unreadable] [unreadable] 3. Characterization of mammalian mediator.[unreadable] [unreadable] 4. Reconstitution of yeast TAF-dependent transcription in vitro.[unreadable] [unreadable] 5. Studies of RSC-nucleosome interaction.